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    Biocontrol Potential, Genome and Nonribosomal Peptide Synthetase Gene Expression of Bacillus velezensis 2211
    (2023-01-01)
    Niemhom, Nantawan
    ;
    Members of the genus Bacillus produced a diverse group of antimicrobial compounds. Here, we presented the antifungal activity and genome sequence analysis of Bacillus sp. 2211, a potential plant-growth-promoting bacterium. Bacterial supernatants from strain 2211 cultures in nutrient broth (NB) and potato dextrose broth (PDB) suppressed the mycelial growth of Pyricularia oryzae, Colletotrichum aenigma, Colletotrichum fructicola and Fusarium oxysporum. The supernatants were also able to suppress spore germination of these fungi, except for F. oxysporum. However, the supernatant from PDB displayed a significantly higher inhibition activity than NB. Additionally, the supernatant from PDB significantly reduced the disease severity caused by P. oryzae on rice seedlings. The genome of strain 2211 was sequenced. The highest digital DNA-DNA hybridization (80.1%) and average nucleotide identity (97.57%) levels indicated that strain 2211 was a member of the species Bacillus velezensis. The phylogenomic analysis showed that it clustered with B. velezensis NRRL B-41580<sup>T</sup>, B. velezensis KACC 13105 and B. velezensis subsp. plantarum FZB42<sup>T</sup>. The gene expression analysis showed the up-regulation of nonribosomal peptide synthetase (NRPS) genes bmyA, fenB and dhbE in PDB, compared to NB. This work demonstrated that the culture media affected the antagonistic activity of strain 2211 possibly through the modification of NRPS biosynthesis genes.
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    Evaluations of the mutagenicity of a pigment extract from bulb culture of Hippeastrum reticulatum
    (2014-01-01)
    Nitteranon, Viriya
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    Vuttipongchaikij, Supachai
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    Sakulkoo, Jenjira
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    Srijakkoat, Monthira
    The use of anthocyanins in food products as colorants has been limited because of their instability toward alkaline pH and high temperature. This study aimed to determine color stability and mutagenicity of the anthocyanin-based pigment extract from bulb cultures of Hippeastrum (Hippeastrum reticulatum). The pigment extract retained its reddish-orange color under alkaline conditions (≤pH 11) and was stable up to 6h at 95°C. The mutagenicity of the extract was evaluated in vitro and in vivo. Hippeastrum pigment extract up to 1.25mgplate<sup>-1</sup> was found non-mutagenic in Ames test using Salmonella typhimurium strain TA98 and TA100. Chromosome aberrations were observed when human lymphocytes were treated with the extract up to 1.5mgml<sup>-1</sup>. However, the extract up to 1.4mgml<sup>-1</sup> was found to exhibit relatively low or no mutagenicity in in vitro comet assays with human lymphocytes. In in vivo micronucleated reticulocyte assay, mice were treated orally with the extract up to 1gkg<sup>-1</sup>. No significant increase of the percentage of micronucleated peripheral reticulocytes compared to the negative control groups was found. Taken together, our study indicates that Hippeastrum pigment extract is potentially applicable as an additive colorant in the diet and related products. © 2014 Elsevier Ltd.
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    Paenibacillus lemnae sp. nov., an endophytic bacterium of duckweed (Lemna aequinoctialis)
    A Gram-stain-variable, rod-shaped and endospore-forming bacterium, designated strain L7-75, was isolated from duckweed (Lemna aequinoctialis). Cells were motile with a monopolar flagellum. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain L7-75<sup>T</sup> belonged to the genusPaenibacillus, and the closest phylogenetically related species werePaenibacillus uliginis N3/975<sup>T</sup> (98.5 % 16S rRNA gene sequence similarity),Paenibacillus purispatii ES_M17<sup>T</sup> (98.5 %), Paenibacillus lactis MB 1871<sup>T</sup> (98.2 %), Paenibacillus campinasensis 324<sup>T</sup> (97.7 %), Paenibacillus glucanolyticusS93<sup>T</sup> (97.7 %) and Paenibacillus lautus ATCC 43898<sup>T</sup> (97.4 %). Growth of strain L7-75<sup>T</sup> was observed at pH 7–10 and at 20–40 °C, and NaCl concentrations up to 5 % (w/v) were tolerated. Major cellular fatty acids included anteiso-C<inf>15 : 0</inf>, C<inf>16 : 0</inf> and anteiso-C<inf>17 : 0</inf> that were present at 36.0 %, 14.2 % and 10.0 % of the total cellular fatty acid profile, respectively. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidyl-N-methylethanolamine. MK-7 was the predominant menaquinone. The diamino acid found in the cell-wall peptidoglycan wasmeso-diaminopimelic acid. The DNA G+C content was 49.1 mol% (T<inf>m</inf>). DNA–DNA relatedness values between strain L7-75<sup>T</sup> and its closest relatives ranged from 4.4 to 47.8 %. These results indicate that strain L7-75<sup>T</sup> represents a novel species of the genus Paenibacillus, for which the name Paenibacillus lemnaesp. nov. is proposed. The type strain is L7-75<sup>T</sup> ( = BCC 67838<sup>T</sup> = NBRC 109972<sup>T</sup>).
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    Pseudoxanthomonas helianthi sp. Nov., isolated from roots of Jerusalem artichoke (Helianthus tuberosus)
    A bacterium designated as strain roo10<sup>T</sup> was isolated from roots of Jerusalem artichoke (Helianthus tuberosus). Cells were Gram-stain-negative and non-motile rods. The phylogenetic analysis of the 16S rRNA gene indicated that it represented a member of the genus Pseudoxanthomonas, and its close relatives included Pseudoxanthomonas kalamensis JA40<sup>T</sup> (97.8% 16S rRNA gene sequence similarity), Pseudoxanthomonas sangjuensis 5GH38-5<sup>T</sup> (97.7%) and Pseudoxanthomonas daejeonensis TR6-08<sup>T</sup> (97.1%). Growth of roo10<sup>T</sup> occurred at pH 7-9. The temperature for growth ranged from 20 to 37 °C. Tolerance to NaCl was observed from 0.005 to 5% (w/v) concentration. Predominant fatty acids were iso-C<inf>15:0</inf> (23.5%), iso-C<inf>16:0</inf> (18.9%) and anteiso-C<inf>15:0</inf> (11.5%). Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidyl-N-methylethanolamine were the major polar lipids. The predominant quinone was ubiquinone 8 (Q-8). The DNA G+C content was 65.7 mol% [from melting temperature (T<inf>m</inf>)]. Comparison of phenotypic and chemotaxonomic characteristics indicated that roo10<sup>T</sup> was distinguishable from its close relatives. Additionally, the DNA-DNA relatedness levels between roo10<sup>T</sup> and P. kalamensis DSM 18571<sup>T</sup> (22±0.5%), P. sangjuensis 5GH38-5<sup>T</sup> (21±0.2%) and P. daejeonensis DSM 17801<sup>T</sup> (3±1%) were lower than 70%. These results indicated that roo10<sup>T</sup> represented a novel species of the genus Pseudoxanthomonas, for which the name Pseudoxanthomonas helianthi sp. nov. is proposed. The type strain is roo10<sup>T</sup> (=BCC 70700<sup>T</sup>=NBRC 110414<sup>T</sup>).
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    Effects of nutrient media on vegetative growth of Lemna minor and Landoltia punctata during in vitro and ex vitro cultivation
    (2013-03-28) ;
    Vuttipongchaikij, Supachai
    Lemnaceous plants, namely Lemna minor and Landoltia punctata, have been used in various types of biological research. The effects of Murashige and Skoog (MS) and Hoagland media on vegetative growth rate of both species during in vitro and ex vitro cultivation were investigated. Under axenic conditions, frond proliferation of L. minor and Lan. punctata in Hoagland medium are 8 and 11.5% respectively faster than that in MS medium. Biomass production in Hoagland medium also increases 2.2-fold (L. minor) and 1.4- fold (Lan. punctata) compared to MS medium. The roots of both species in MS medium are distinctly shorter than those in Hoagland medium. In contrast, ex vitro regeneration of frond colonies in MS medium is 22.2% (for L. minor) and 17.1% (for Lan. punctata) faster than in Hoagland medium. Similarly, ex vitro biomass production of both species in MS increases 1. 8-fold (for L. minor) and 1.3-fold (for Lan. punctata) compared to that in Hoagland medium. Root elongation of the frond colonies in MS and Hoagland media is comparable. The distinct effects of MS and Hoagland media on vegetative growth of both species and the pre-determination of ex vitro growth rates in each medium are demonstrated. © 2013 by Maejo University, San Sai, Chiang Mai, 50290 Thailand. Reproduction is permitted for noncommercial purposes.
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    Chitinophaga oryzae sp. Nov., an epiphytic bacterium isolated from rice root surfaces
    (2021-01-01)
    Niemhom, Nantawan
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    Suriyachadkun, Chanwit
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    Two Gram-stain-negative, non-motile, rod-shaped bacterial strains were isolated from the surfaces of rice roots. They were designated as strains 1303<sup>T</sup> and 1310. Their colonies were circular, entire, opaque, convex and yellow. They were chitinaseand catalase-positive, reduced nitrate and grew at 16–37 °C (optimum, 30 °C), pH 5.0–10.0 (optimum, pH 7.0) and 0–2.0% NaCl (optimum, 1.0%). Based on the 16S rRNA gene sequence analysis, they were classified as members of the genus Chitinophaga. Results of phylogenetic and phylogenomic analyses indicated that they formed a cluster with Chitinophaga eiseniae YC6729<sup>T</sup>, Chitinophaga qingshengii JN246<sup>T</sup>, Chitinophaga varians 10-7 W-9003<sup>T</sup> and Chitinophaga fulva G-6-1-13<sup>T</sup>. When the genomic sequences of strains 1303<sup>T</sup> and 1310 were compared with their close relatives, the average nucleotide identity and digital DNA– DNA hybridization values were below the cut-off levels. Phosphatidylethanolamine was the major polar lipid. MK-7 was the major respiratory quinone. iso-C<inf>15:0</inf>, C<inf>16:1</inf> ω5c, iso-C<inf>17:0</inf> 3-OH and summed feature 3 (C<inf>16:1</inf> ω7c/C<inf>16:1</inf> ω6c) were the predominant fatty acids. Differential characteristics between both strains and their close relatives were also observed. Based on the distinctions in genotypic, phenotypic and chemotypic features, strains 1303<sup>T</sup> and 1310 represent members of a novel species of the genus Chitinophaga, for which the name Chitinophaga oryzae sp. nov. is proposed. The type strain is 1303<sup>T</sup> (=KACC 22075<sup>T</sup>=TBRC 12926<sup>T</sup>).
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    Light-Emitting Diode Illumination Enhances Biomass, Pigment, and Lipid Production in Halotolerant Cyanobacterium Aphanothece halophytica
    (2025-06-01)
    Thongtha, Sitthichai
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    Incharoensakdi, Aran
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    Light characteristics, including spectrum and intensity, significantly impact cyanobacterial biomass production, pigment biosynthesis, and cellular metabolism, influencing the composition of various biochemical compounds. This study aimed to investigate the effects of light-emitting diode (LED) illumination on biomass, pigment, and lipid production in the unicellular halotolerant cyanobacterium Aphanothece halophytica, cultivated in a suitable natural seawater (SNSW) medium. The results revealed that LED light outperformed fluorescent light, with blue LED light, particularly at an intensity of 60 μmol photons m<sup>−2</sup> s<sup>−1</sup>, significantly enhancing growth, pigment synthesis, and lipid accumulation. This resulted in a maximum cell density of 68.96 ± 1.52 × 10<sup>6</sup> cells mL<sup>−1</sup>, a specific growth rate of 0.302 ± 0.002 day<sup>−1</sup>, and a lipid productivity of 56.81 ± 0.75 mg L<sup>−1</sup> day<sup>−1</sup>. White LED light produced lipids suitable for biodiesel, whereas blue, green, and red LEDs promoted the accumulation of polyunsaturated fatty acids (PUFAs), beneficial for food supplements. These findings highlight the potential of LED-based cultivation strategies for optimizing biomass and biochemical compound production in A. halophytica.
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    Differential effects of synthetic media on long-term growth, starch accumulation and transcription of ADP-glucosepyrophosphorylase subunit genes in Landoltia punctata
    Murashige & Skoog (MS) and Hoagland’s media were previously used for in vitro culture of Landoltia punctata. During subsequent ex vitro culture, the use of MS medium resulted in a higher growth rate, compared to Hoagland’s medium. Thus, a higher starch content of L. punctata in MS medium was previously hypothesized. Here, L. punctata strain 5632 was isolated and characterized using morphological characteristics and the atpF-atpH intergenic region. During early cultivation stage, fresh weight and relative growth rate in MS medium were lower than Hoagland’s medium. Conversely, starch content in MS medium was considerably higher than in Hoagland’s medium. Medium effects on expression of genes coding for starch-biosynthesis ADP-glucosepyrophosphorylase (AGPase) were determined. Genomic fragments of small (LeAPS) and large (LeAPL1) AGPase subunits were characterized. Differential expression between each AGPase subunit genes was observed in both media. Additionally, in MS medium, the highest correlation coefficients between starch content and gene expression was found with LeAPS (0.81) and followed by LeAPL3 (0.67), LeAPL2 (0.65) and LeAPL1 (0.28). In Hoagland’s medium, the coefficients of LeAPL3 (0.83) and LeAPL2 (0.62) were higher than LeAPS (0.18) and LeAPL1 (−0.62). This suggested different levels of contributions of these genes in starch biosynthesis in both media.
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    Biogas effluent treatment by Landoltia punctata for starch biomass production
    (2023-05-01)
    Kirdponpattara, Suchata
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    Phisalaphong, Muenduen
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    Chanroj, Salil
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    Sriariyanun, Malinee
    Due to the high potential of using Landoltia punctata in biogas effluent treatment and starch accumulation, three integrated processes were proposed: (1) bioethanol production from duckweed, (2) biogas production from vinasse, and (3) biogas effluent treatment with sequentially increasing starch-rich duckweed biomass. This recent work focuses only on the third process, which involves the treatment of wastewater from biogas effluent and the production of L. punctata with high starch content. L. punctata showed a significantly higher TKN removal and biomass production than Lemna aequinoctialis. After nine days of wastewater treatment and five days of starch accumulation, L. punctata grew by 4.3 g/m<sup>2</sup>/day, with 44.4% starch content. NH<inf>3</inf>, NH<inf>4</inf><sup>+</sup>, and PO<inf>4</inf><sup>3−</sup> in the biogas effluent were rapidly utilized in the first three days for L. punctata's growth. The upregulation of AGPase genes in the starch biosynthesis pathway was observed when the environmental conditions changed in both nutrient starvation and population reduction. Production of starch-rich L. punctata using biogas effluent is a sustainable feedstock for the bioethanol process. Moreover, L. punctata-based stabilization pond is a promising technique for biogas effluent treatment.
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    Micromonospora endophytica sp. nov., an endophytic actinobacteria of Thai upland rice (Oryza sativa)
    An actinobacterial strain, DCWR9-8-2 T, was isolated from a leaf of Thai upland rice (Oryza sativa) collected in Chumporn province, Thailand. Strain DCWR9-8-2 T is Gram-stain-positive aerobic bacteria that produce single spores directly on the vegetative hypha. Cell wall peptidoglycan of this strain exhibits meso-diaminopimelic acid and glycine, the reducing sugars of whole-cell hydrolysate are arabinose, glucose, ribose, xylose and small amount of mannose. The phospholipid profiles in the membrane are comprised of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides. The major menaquinones are MK-9(H 4) and MK-10(H 6). The diagnostic cellular fatty acids are iso-C 16:0 and iso-C 15:0. The G+C content of the genomic DNA is 72.5 mol%. The result of 16S rRNA sequence analysis of the strain revealed that this strain was closely related to Micromonospora auratinigra TT1-11 T (99.25%). On the other hand, the result of gyrB gene sequence analysis revealed that this strain was closed to M. eburnea JCM 12345 T (96.30%). In addition, a combination of DNA-DNA hybridization results and some phenotypic properties supported that this strain should be judged as a novel species of the genus Micromonospora, for which the name M. endophytica sp. nov. is proposed. The type strain is DCWR9-8-2 T (=BCC 67267 T =NBRC 110008 T).
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